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      Development of ultraviolet spectroscopic method for the estimation of metronidazole benzoate from pharmaceutical formulation

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          Abstract

          Background:

          The present study was undertaken with an objective to develop a simple, accurate, cost-effective and reproducible ultraviolet spectrophotometric method for the estimation of metronidazole benzoate (MB) from pharmaceutical formulations.

          Materials and Methods:

          The analysis was performed on λ max 268 nm by using 0.1 NHCl as diluents. The proposed method was validated on International Conference Harmonization guideline including the parameters viz., accuracy, linearity, precision, specificity and reproducibility. The proposed method was also used to access the content of MB in two commercial brands of Indian market.

          Results:

          Beer's law was obeyed in the concentration range of 1-10 μg/ml having regression equation y = 0.078 x-0.012. The accuracy value for 4 μg/ml and 5 μg/ml concentration of MB was found to be 99.37% and 98.9% respectively. The relative standard deviation of interday and intraday was lesser than 1%. The developed method was applied on two different marketed brands and contents of MB were found to be 98.62% and 98.59% incompliance with labeled claim. The results were under the limit of acceptance statistically.

          Conclusion:

          It was concluded that the proposed method can be used for routine analysis of MB in bulk and commercial formulations.

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          Most cited references12

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          Method validation and determinations of levofloxacin, metronidazole and sulfamethoxazole in an aqueous pharmaceutical, urine and blood plasma samples using quantitative nuclear magnetic resonance spectrometry.

          Selective, rapid and accurate quantitative proton nuclear magnetic resonance (qHNMR) method for the determination of levofloxacin, metronidazole benzoate and sulfamethoxazole in aqueous solutions was developed and validated. The method was successfully applied to the determinations of the drugs and their admixtures in pharmaceutical, urine and plasma samples. Maleic acid and sodium malate were used as internal standards. Effect of temperature on spectral measurements was evaluated. Linear dynamic ranges of 0.50-68.00, 0.13-11.30 and 0.24-21.00 mg per 0.60 mL solution were obtained for levofloxacin, metronidazole benzoate and sulfamethoxazole, respectively. Average recovery % in the range of 96.00-104.20 ± (0.17-2.91) was obtained for drugs in pure, pharmaceutical, plasma and urine samples. Inter and intra-day analyses gave average recoveries % in the ranges 96.10-98.40 ± (1.68-2.81) and 96.00-104.20 ± (0.17-2.91), respectively. Instrumental detection limits ≤0.03 mg per 0.6 mL were obtained for the three drugs. Developed method has demonstrated high performance characteristics for analyzing investigated drugs and their admixtures. Student t-test at 95% confidence level revealed insignificant bias between the real and measured contents of investigated drugs in pure, pharmaceutical, urine and plasma samples and its admixtures. Application of the statistical F-test revealed insignificant differences in precisions between the developed method and arbitrary selected reference methods.
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            A stability-indicating HPLC assay for metronidazole benzoate.

            A simple and rapid stability-indicating HPLC assay procedure has been developed and validated for metronidazole benzoate. The HPLC conditions were as follows, column: Waters Symmetry C8, 5 microm packing, 4.6 mm x 250 mm; detection: UV at 271 nm; injection volume: 20 microl; mobile phase: acetonitrile-0.1% glacial acetic acid in monobasic potassium phosphate (0.01 M) (40:60, v/v); isocratic elution under ambient temperature at 2.0 ml min(-1). The procedure separated metronidazole benzoate and its potential degradation products, metronidazole and benzoic acid, in an overall analysis time of about 6 min with metronidazole benzoate eluting at about 5 min. The injection repeatability was 0.03%, and the intraday and interday repeatability were 0.4 and 0.7%, respectively. The procedure provided a linear response over the concentration range 0.2-800 microg ml(-1) (r=1.0000) with the limits of detection and quantitation 0.03 and 0.2 microg ml(-1), respectively. The solubilities of metronidazole benzoate in water, 0.01 M hydrochloric acid and 0.05 M phosphate buffer, pH 6.8, determined each in triplicate using the procedure, were 0.2 mg ml(-1) (R.S.D. 7%), 0.4 mg ml(-1) (R.S.D. 2%) and 0.2 mg ml(-1) (R.S.D. 8%), respectively. The results show no detectable hydrolysis of metronidazole benzoate in 0.01 M hydrochloric acid at 37 degrees C or in the mobile phase at ambient temperature in 10 h.
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              Simultaneous determination of ranitidine and metronidazole in human plasma using high performance liquid chromatography with diode array detection.

              The development and validation of a simple method for the simultaneous determination of ranitidine and metronidazole in human plasma is described. Plasma samples (250 microL) were deproteinized by precipitation with 60% perchloric acid, centrifuged and the supernatant directly injected into the HPLC. Separation was achieved in isocratic mode with a Shimpak C(18) column and a mobile phase consisting of 10mM potassium dihydrogen phosphate pH 3.5:acetonitrile (90:10, v/v) with UV detection at 315 nm. The method showed good selectivity and sensitivity. Good and consistent recovery for metronidazole and ranitidine was obtained: 96.22+/-3.52 and 95.00+/-4.50% for ranitidine (25-1000 ng/mL) and metronidazole (60-10,000 ng/mL), respectively (n=3). With this one-step sample preparation method, both ranitidine and metronidazole could be quantified simultaneously in human plasma with good precision (R.S.D.<15%) and accuracy (bias values below 15%). The limit of quantification for ranitidine and metronidazole were 20 and 40 ng/mL plasma, respectively.
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                Author and article information

                Journal
                J Nat Sci Biol Med
                J Nat Sci Biol Med
                JNSBM
                Journal of Natural Science, Biology, and Medicine
                Medknow Publications & Media Pvt Ltd (India )
                0976-9668
                2229-7707
                Jul-Dec 2014
                : 5
                : 2
                : 261-264
                Affiliations
                [1] Department of Pharmaceutical Chemistry, Central Facility of Instrumentation, School of Pharmaceutical Sciences, IFTM University, Moradabad, Uttar Pradesh, India
                Author notes
                Address for correspondence: Dr. Arun K. Mishra, Central Facility of Instrumentation, School of Pharmaceutical Sciences, IFTM University, Moradabad - 244 001, Uttar Pradesh, India. E-mail: arun_azam@ 123456rediffmail.com
                Article
                JNSBM-5-261
                10.4103/0976-9668.136154
                4121894
                d3179352-2566-49c4-9771-60a0147d6a11
                Copyright: © Journal of Natural Science, Biology and Medicine

                This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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                Categories
                Original Article

                Life sciences
                international conference harmonization guidelines,metronidazole benzoate,quality control,ultraviolet spectrometry

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